Selective sweep analysis of the adaptability of the Yarkand hare (Lepus yarkandensis) to hot arid environments using SLAF-seq.

The Yarkand hare (Lepus yarkandensis) inhabits arid desert areas and is endemic to China. It has evolved various adaptations to survive in hot arid environments, including stress responses, the ability to maintain water homeostasis and heat tolerance. Here, we performed a selective sweep analysis to identify the candidate genes for adaptation to hot arid environments in the Yarkand hare. A total of 397 237 single-nucleotide polymorphisms were obtained from 80 Yarkand hares, which inhabit hot arid environments, and 36 Tolai hares (Lepus tolai), which inhabit environments with a mild climate, via specific-locus amplified fragment sequencing. We identified several candidate genes that were associated with the heat stress response (HSPE1), oxidative stress response (SLC23A and GLRX2), immune response (IL1R1 and IRG1), central nervous system development (FGF13, THOC2, FMR1 and MECP2) and regulation of water homeostasis (CDK1) according to fixation index values and θπ ratios in the selective sweep analysis, and six of these genes (GLRX2, IRG1, FGF13, FMR1, MECP2 and CDK1) are newly discovered genes. To the best of our knowledge, this is the first study to identify candidate genes for adaptation to hot arid environments in the Yarkand hare. The results of this study enhance our understanding of the adaptation of the Yarkand hare to hot arid environments and will aid future studies aiming to functionally verify these candidate genes.

Airborne signals of Pseudomonas fluorescens modulate swimming motility and biofilm formation of Listeria monocytogenes in a contactless coculture system.

Bacterial volatile compounds (BVCs) facilitate interspecies communication in socio-microbiology across physical barriers, thereby influencing interactions between diverse species. The impact of BVCs emitted from Pseudomonas on the biofilm formation characteristics of Listeria monocytogenes within the same ecological niche has been scarcely investigated under practical conditions of food processing. The objective of this study was to explore the motility and biofilm formation characteristics of L. monocytogenes under the impact of Pseudomonas BVCs. It was revealed that BVCs of P. fluorescens, P. lundensis, and P. fragi significantly promoted swimming motility of L. monocytogenes (P < 0.05). As evidenced by crystal violet staining, the L. monocytogenes biofilms reached a maximum OD570 value of approximately 3.78 at 4 d, which was 0.65 units markedly higher than that of the control group (P < 0.05). Despite a decrease in adherent cells of L. monocytogenes biofilms among the BVCs groups, there was a remarkable increase in the abundance of extracellular polysaccharides and proteins with 3.58 and 4.90 µg/cm2, respectively (P < 0.05), contributing to more compact matrix architectures, which suggested that the BVCs of P. fluorescens enhanced L. monocytogenes biofilm formation through promoting the secretion of extracellular polymers. Moreover, the prominent up-regulated expression of virulence genes further revealed the positive regulation of L. monocytogenes under the influence of BVCs. Additionally, the presence of BVCs significantly elevated the pH and TVB-N levels in both the swimming medium and biofilm broth, thereby exhibiting a strong positive correlation with increased motility and biofilm formation of L. monocytogenes. It highlighted the crucial signaling regulatory role of BVCs in bacterial interactions, while also emphasizing the potential food safety risk associated with the hitchhiking behavior of L. monocytogenes, thereby shedding light on advancements in control strategies for food processing.

The role of PhoP/PhoQ system in regulating stress adaptation response in Escherichia coli O157:H7.

The development of acid tolerance response (ATR) as a result of low pH in Escherichia coli O157:H7 (E. coli O157:H7) contaminating beef during processing is considered a major food safety concern. Thus, in order to explore the formation and molecular mechanisms of the tolerance response of E. coli O157:H7 in a simulated beef processing environment, the resistance of a wild-type (WT) strain and its corresponding ΔphoP mutant to acid, heat, and osmotic pressure was evaluated. Strains were pre-adapted under different conditions of pH (5.4 and 7.0), temperature (37 °C and 10 °C), and culture medium (meat extract and Luria-Bertani broth media). In addition, the expression of genes related to stress response and virulence was also investigated among WT and ΔphoP strains under the tested conditions. Pre-acid adaptation increased the resistance of E. coli O157:H7 to acid and heat treatment while resistance to osmotic pressure decreased. Moreover, acid adaptation in meat extract medium simulating slaughter environment increased ATR, whereas pre-adaptation at 10 °C reduced the ATR. Furthermore, it was shown that mildly acidic conditions (pH = 5.4) and the PhoP/PhoQ two-component system (TCS) acted synergistically to enhance acid and heat tolerance in E. coli O157:H7. Additionally, the expression of genes related to arginine and lysine metabolism, heat shock, and invasiveness was up-regulated, which revealed that the mechanism of acid resistance and cross-protection under mildly acidic conditions was mediated by the PhoP/PhoQ TCS. Both acid adaptation and phoP gene knockout reduced the relative expression of stx1 and stx2 genes which were considered as critical pathogenic factors. Collectively, the current findings indicated that ATR could occur in E. coli O157:H7 during beef processing. Thus, there is an increased food safety risk due to the persistence of tolerance response in the following processing conditions. The present study provides a more comprehensive basis for the effective application of hurdle technology in beef processing.

Potential inhibitory effect of carbon dioxide on the spoilage behaviors of Pseudomonas fragi in high-oxygen packaged beef during refrigerated storage.

Pseudomonas fragi is a dominant meat spoilage organism under high-oxygen modified atmosphere packaging (HiOx-MAP). This work investigated the effects of CO2 on P. fragi growth and the related spoilage phenomena of HiOx-MAP beef. Minced beef incubated with P. fragi T1, a strain owning the strongest spoilage potential among isolates, was stored under CO2-enriched HiOx-MAP (TMAP; 50% O2/40% CO2/10% N2) or non-CO2 HiOx-MAP (CMAP; 50% O2/50% N2) at 4 °C for 14 days. Compared to CMAP, TMAP maintained sufficient O2 levels to endow beef with higher a* values and meat color stability due to lower P. fragi counts from day 1 (P < 0.05). TMAP samples also showed lower (P < 0.05) lipase activity and protease activity within 14-days and 6-days than CMAP samples respectively. TMAP delayed the significantly increased pH and total volatile basic nitrogen contents occurred in CMAP beef during storage. Despite TMAP markedly promoted the lipid oxidation associated with higher concentrations of hexanal and 2,3-octanedione than CMAP (P < 0.05), TMAP beef retained an acceptable organoleptic odor due to a CO2-inhibition on the microbial-induced 2,3-butanedione and ethyl 2-butenoate formation. This study provided a comprehensive insight into the antibacterial mechanism of CO2 on P. fragi in HiOx-MAP beef.

Processing interventions for enhanced microbiological safety of beef carcasses and beef products: A review.

Chilled beef is inevitably contaminated with microorganisms, starting from the very beginning of the slaughter line. A lot of studies have aimed to improve meat safety and extend the shelf life of chilled beef, of which some have focused on improving the decontamination effects using traditional decontamination interventions, and others have investigated newer technologies and methods, that offer greater energy efficiency, lower environmental impacts, and better assurances for the decontamination of beef carcasses and cuts. To inform industry, there is an urgent need to review these interventions, analyze the merits and demerits of each technology, and provide insight into 'best practice' to preserve microbial safety and beef quality. In this review, the strategies and procedures used to inhibit the growth of microorganisms on beef, from slaughter to storage, have been critiqued. Critical aspects, where there is a lack of data, have been highlighted to help guide future research. It is also acknowledge that different intervention programs for microbiological safety have different applications, dependent on the initial microbial load, the type of infrastructures, and different stages of beef processing.

The acid tolerance responses of the Salmonella strains isolated from beef processing plants.

The development of the stationary-phase, low-pH-inducible acid tolerance response (ATR) in the Salmonella contaminant of beef during the processing arises food safety concerns, because it may evoke bacterial coping mechanisms against bactericidal insults and alter gene expression that contribute to pathogen virulence. However, information on the development of the ATR and the stability (defined as the capacity to maintain the acquired acid tolerance after induction) in the Salmonella during the production and distribution of beef is limited. After adaptation overnight, ATRs in the 79 strains of Salmonella isolated from beef processing plants were investigated by comparing the log reduction in the 2-h acid challenge trials at pH 3.0. Six representative strains were selected to further estimate the effect of three factors in the incubation period on the development of the ATR, including adapted pH values (5.0, 5.4, 6.0, and 7.0), temperatures (10 °C and 37 °C), and the adaptation media (meat extract and brain heart infusion media). The stability of acid tolerance during the long-time chilled storage (4 °C for 13 days) was also observed on two strains of serotypes S. Derby and S. Meleagridis. All the strains isolated from beef processing plants exhibited an enhanced acid tolerance indicating the widespread existence of ATR. The results also revealed that strain variability was present in the development of ATR. Significant tolerance to lethal acidic environments (pH 3.0) was found when the Salmonella strains had been acid-adapted in meat extract at pH 5.0, pH 5.4, or pH 6.0, which indicated the possible induction of ATR during beef production. After the acid adaptations, the population reduction after the acid challenge (BHI, pH = 3) in the strains was significantly lower than the non-induced at the 1d, 7 day and 13 day's storage in meat extract media at 4 °C, which revealed the persistence of ATR during beef distribution. Compared to 37 °C, adaptation in lower temperature (10 °C) significantly reduced the ATR and no ATR was developed when adapted in 4 °C. This emphasizes the importance of keeping a low temperature of beef throughout the supply chains of beef industry.

Remediation of copper-contaminated soils using Tagetes patula L., earthworms and arbuscular mycorrhizal fungi.

Arbuscular mycorrhizal fungi (AMF) and earthworms have potential uses in the bioremediation of contaminated soils. In recent years, heavy metal-contaminated sites have been remediated by adding plants and AMF or earthworms to the soil. However, there are few studies on remediation using combinations of plants, animals, and microbes, especially for the remediation of Cu-contaminated soil. The present study investigated the separate and combined effects of AMF and earthworms on Cu-contaminated soil in which Tagetes patula L. was grown. The results show that the combined application of AMF and earthworms markedly increased the biomass of plant shoots and roots by more than 100%. It also increased Cu extraction by T. patula by 270%. The combined treatment was effective in increasing the CEC, contents of OM, and available Cu, P and K, but reduced the soil pH. Furthermore, the combined treatment significantly increased the abundance and diversity of the soil microbial community. In particular, the abundances of the bacteria Bacteroides, Proteobacteria, and Actinobacteria were increased, with the genera Flavobacterium, Pedobacter, Algoriphagus, Gaetbulibacter, Pseudomonas, Luteimonas, and Arthrobacter dominating. Meanwhile, the abundance of the fungus Zygomycota was increased, with Mortierella dominating. Moreover, inoculation with earthworms greatly improved the structure of the soil microbial community.

Compared with phytoremediation or microbial remediation alone, combined remediation was considered to be a more efficient remediation strategy. This study describes the effects of the combined remediation of T. patula, AMF, and earthworms on the physiochemical properties and microbial community of Cu-contaminated soil. This provides a comprehensive strategy to improve the phytoremediation efficiency in Cu-contaminated soil.

Sensitivity of the Transport of Plastic Nanoparticles to Typical Phosphates Associated with Ionic Strength and Solution pH.

The influence of phosphates on the transport of plastic particles in porous media is environmentally relevant due to their ubiquitous coexistence in the subsurface environment. This study investigated the transport of plastic nanoparticles (PNPs) via column experiments, paired with Derjaguin-Landau-Verwey-Overbeek calculations and numerical simulations. The trends of PNP transport vary with increasing concentrations of NaH2PO4 and Na2HPO4 due to the coupled effects of increased electrostatic repulsion, the competition for retention sites, and the compression of the double layer. Higher pH tends to increase PNP transport due to the enhanced deprotonation of surfaces. The release of retained PNPs under reduced IS and increased pH is limited because most of the PNPs were irreversibly captured in deep primary minima. The presence of physicochemical heterogeneities on solid surfaces can reduce PNP transport and increase the sensitivity of the transport to IS. Furthermore, variations in the hydrogen bonding when the two phosphates act as proton donors will result in different influences on PNP transport at the same IS. This study highlights the sensitivity of PNP transport to phosphates associated with the solution chemistries (e.g., IS and pH) and is helpful for better understanding the fate of PNPs and other colloidal contaminants in the subsurface environment.

Acid tolerance response of Salmonella during simulated chilled beef storage and its regulatory mechanism based on the PhoP/Q system.

To investigate the persistence of acid tolerance response (ATR) and the regulatory mechanism during chilled storage, Salmonella ATCC 14028 and the △phoP mutant were acid adapted and then incubated in meat extract at 4 °C for 24 days as simulated beef storage. The bacterial population, D values and expression of PhoP/PhoQ linked genes of both strains were determined at 6-day intervals. Although a mild suppression effect on the D values of adapted Salmonella was found during the long-time storage in meat extract at 4 °C, the D value of adapted strains was significantly higher than non-adapted strains, indicating the persistence of ATR during the whole aging and distribution of beef posing a threat to food safety. The fact that low temperature inhibits the formation of ATR at the early adapted stage emphasizes the importance of keeping a low-temperature environment during slaughter. An interaction between the acidic adaptation and phoP gene on D values was found and the expression levels of adiA, adiY, cadA and cadB genes was significantly reduced in the △phoP mutant, suggesting that PhoP/Q system plays an important role in the ATR by sensing the pH and regulating lysine and arginine decarboxylation directly or indirectly.

Lactic Acid and Peroxyacetic Acid Inhibit Biofilm of Escherichia coli O157:H7 Formed in Beef Extract.

The objective of this study was to evaluate the inhibitory effect of lactic acid (LA) and peroxyacetic acid (PAA) on the biofilm formation of Escherichia coli O157:H7 in beef extract (BE). BE medium was used as the growth substrate in this study, to make the control effect closer to the situation of the factory. The biofilm inhibitory efficacy of LA and PAA was tested by using a crystal violet staining assay and microscopic examination. And then, extracellular polymeric substance (EPS) production, metabolic activity, and real-time polymerase chain reaction assay were used to reveal the biofilm inhibition mechanism of LA and PAA. The results showed that both LA and PAA significantly inhibited biofilm formation of E. coli O157:H7 at minimum inhibitory concentrations (MICs) (p < 0.05). At MIC, LA and PAA showed different effects on the biofilm metabolic activity and the EPS production of E. coli O157:H7. Supporting these findings, expression analysis showed that LA significantly suppressed quorum sensing genes (luxS and sdiA) and adhesion genes (flhC), while PAA downregulated the transcription of extracellular polysaccharide synthesis genes (adrB and adrA) and the global regulatory factor csgD. This result revealed that LA and PAA had different biofilm inhibitory mechanisms on E. coli O157:H7; LA inhibited the biofilm formation mainly by inhibiting metabolic activity, while PAA inhibited EPS production. This study provided a theoretical basis for the control of E. coli O157:H7 biofilm in the actual production process.

Comparative transcriptomic analysis reveals novel insights into the response to Cr(VI) exposure in Cr(VI) tolerant ectomycorrhizal fungi Pisolithus sp. 1 LS-2017.

Chromium (Cr) is one of the most toxic heavy metals and a health hazard to millions of people worldwide. Ectomycorrhizal (ECM) fungi can assist plants in phytoremediation of heavy metal contaminated soil. Cr tolerance differs among ECM fungal varieties, but the underlying molecular mechanisms of Cr tolerance in ECM fungi are not clear. This study identified, analysed and compared the Cr(VI)-induced transcriptional changes between Cr(VI)-tolerant strain (Pisolithus sp. 1 LS-2017) and Cr(VI)-sensitive strain (Pisolithus sp. 2 LS-2017) by de novo transcriptomic analysis. The results showed that 93,642 assembled unique transcripts representing the 22,353 (46.76%) unigenes matched the proteins we have known in the Nr database and 47,801 unigenes were got from the Pisolithus spp. For DEGs between the control and 10 mg/L Cr(VI) treatment, cyanoamino acid metabolic, type I diabetes mellitus metabolism, nitrogen metabolism and beta-Alanine metabolism pathways were significantly enriched (p < 0.05) in Pisolithus sp. 1 LS-2017. Two nitrate reductase family genes (nidD, niiA) provide Cr(VI) tolerance for Pisolithus sp. 1 LS-2017 by regulating Cr(VI) reduction. In addition, NO produced by nidD, niiA regulated denitrification can alleviate Cr(VI) induced oxidative stress. In Pisolithus sp. 2 LS-2017, the alcC, aldA and lcf2 gene may alleviate Cr(VI) induced oxidative stress by protecting SH groups and increasing secondary metabolism, reducing detoxify aldehydes to carboxylic acids and producing LCPUFAs respectively; .T gene regulate Cr(VI) induced wound healing by pigmentation and stability of melanin in spore; MKP2 gene accelerate Cr(VI) induced cell death and gpmA gene regulated Cr(VI) induced energy emergency.

Acid Tolerance Response of Listeria monocytogenes in Various External pHs with Different Concentrations of Lactic Acid.

This study evaluated the acid tolerance response (ATR) of two strains of Listeria monocytogenes (serotype 1/2a and 4b) and one strain of Listeria innocua under different mildly acid conditions. Cells were incubated in combinations of three concentrations of lactic acid medium (3, 4.75, and 15 mM) and three external pH's (pHex 5.0, 6.0, and 6.5), plus, a HCl control, and a blank control (pH 7.4). Results showed that lactic acid induced lower log reduction of all three strains when challenged in severe acid conditions (pH 3.0) after being habituated at a pHex of 5.5 or 6.0 until the log phase, compared with a pHex of 6.5 or the two controls. This indicates that when the pHex was either 5.5 or 6.0 this induced a higher ATR of the strains, which may be caused by the ability of the strains to retain intracellular pH (pHi) homeostasis with pHi maintained in the range of 7.4-7.9. It was also found that a pHex of 5.5 resulted in the highest pHi of the strains across all incubated conditions, which indicates that the pHi may play an important role in the induction of ATR when Listeria cells are habituated in lactic acid, and if the higher pHi can be maintained, the ATR would be stronger. The concentration of lactic acid per se has no significant effect on ATR, which it is proposed was due to the pHi homeostasis maintained within the cells. However, the difference in ATR among three strains was also significant, which cannot be explained by the stable pHi of all tested strains. Therefore, other underlying mechanisms to mediate ATR under different conditions need to be explored in further studies.

Investigation of muscle-specific beef color stability at different ultimate pHs.

OBJECTIVE: This study was aimed to investigate the muscle-specific beef color stability at normal and high ultimate pHs. METHODS: The impact of muscle (Longissimus lumborum [LL] vs psoas major [PM]) and pH (normal ultimate pH [Np] vs high pH dark cutting beef [Hp]) on color stability, indicated by basic color traits, metmyoglobin reducing activity (MRA) and oxygen consumption (OC), as well as the lipid oxidation, were determined over 7 days of display at 4°C. RESULTS: Hp-LL had the highest pH (6.92), followed by Hp-PM (6.01), Np-PM (5.76), and Np-LL (5.52). Hp-LL had increased (p<0.05) a*, chroma and % oxymyoglobin during display. Hp-LL also had the highest metmyoglobin (MMb) reducing activity and OC among all the samples, thus, the greatest color stability, although very dark throughout storage, with lowest values for lightness (L*) and yellowness (b*). Np-LL also exhibited relatively high color stability, as a result of its lower % MMb and OC and higher MRA than psoas muscle samples. The 0.2 unit difference of the pH between Hp and Np psoas muscle, resulted in the difference of the color intensity, not the color stability. Interestingly, high pH psoas muscle (Hp-PM) did not have better color stability than Np-PM, and in fact had lower color stability than even Np-LL. The similar level of OC and lipid oxidation cannot explain the difference in color stability between Hp-PM and Np-LL. CONCLUSION: The Hp does not always show better color stability compared with Np beef, which depends on the muscle type. The balance of MRA and OC is important to keep the color in great intensity and stability in the meantime.

Effect of different short-term high ambient temperature on chicken meat quality and ultra-structure.

OBJECTIVE: This study investigated the effect of different acute heat stress (HS) levels on chicken meat quality and ultra-structure. METHODS: Chickens were randomly divided into 7 groups to receive different HS treatments: i) 36°C for 1 h, ii) 36°C for 2 h, iii) 38°C for 1 h, iv) 38°C for 2 h, v) 40°C for 1 h, vi) 40°C for 2 h, and vii) un-stressed control group (25°C). Blood cortisol level, breasts initial temperature, color, pH, water holding capacity (WHC), protein solubility and ultra-structure were analyzed. RESULTS: HS temperatures had significant effects on breast meat temperature, lightness (L*), redness (a*), cooking loss and protein solubility (p<0.05). The HS at 36°C increased L*24 h value (p<0.01) and increased the cooking loss (p<0.05), but decreased a*24 h value (p<0.05). However, as the temperature increased to 38°C and 40°C, all the values of L*24 h, cooking loss and protein denaturation level decreased, and the differences disappeared compared to control group (p> 0.05). Only the ultimate pH24 h at 40°C decreased compared to the control group (p<0.01). The pH in 36°C group declined greater than other heat-stressed group in the first hour postmortem, which contributed breast muscle protein degeneration combining with high body temperature, and these variations reflected on poor meat quality parameters. The muscle fiber integrity level in group 40°C was much better than those in 36°C with the denatured position mainly focused on the interval of muscle fibers which probably contributes WHC and light reflection. CONCLUSION: HS at higher temperature (above 38°C) before slaughter did not always lead to more pale and lower WHC breast meat. Breast meat quality parameters had a regression trend as HS temperature raised from 36°C. The interval of muscle fibers at 24 h postmortem and greater pH decline rate with high body temperature in early postmortem period could be a reasonable explanation for the variation of meat quality parameters.

Hydrogen ions and organic acids secreted by ectomycorrhizal fungi, Pisolithus sp1, are involved in the efficient removal of hexavalent chromium from waste water.

Pisolithus sp1 is an ectomycorrhizal (ECM) fungi that was chosen during a screening test of six strains of ECM fungi due to its ability to tolerate and remove hexavalent chromium (Cr(VI)). The physiological responses of Pisolithus sp1 to Cr(VI) exposure, the relationship between Pisolithus sp1 and exogenously added organic acids (EAOAs) or Na3VO4 (H+-ATPase inhibitor) and the ability of Pisolithus sp1 to reduce Cr(VI) in liquid culture were also investigated. Hydrogen ions (H+), which were produced directly by Pisolithus sp1, reduced the pH of the medium and played an important role in Cr(VI) reduction; however, Na3VO4 significantly inhibited this process and resulted in a decrease in the Cr(VI) reduction rates. Organic acids were secreted after the reduction in Cr(VI) by Pisolithus sp1, and EAOAs did not significantly affect Cr(VI) reduction; those results revealed the secondary role of organic acids in Cr(VI) reduction. The Cr(VI) removal rate of Pisolithus sp1 approached 99% after Cr(VI) treatment for 12 days. Overall, 75% of the Cr(VI) removal was due to extracellular reduction and 24% was due to adsorption. The results of this study provide a strong basis for using Cr(VI)-tolerant and Cr(VI)-reducing fungi, as well as ectomycorrhiza, in the remediation of Cr(VI)-contaminated sites.

The Characterization of Biofilm Formation and Detection of Biofilm-Related Genes in Salmonella Isolated from Beef Processing Plants.

The biofilm formation behavior of Salmonella isolated from beef processing plants was investigated under varying temperatures (4°C, 10°C, 25°C, 37°C, and 42°C) and pH (4.5, 5.0, 5.5, 6.0, 7.0, and 8.0). The relationships between the presence of biofilm-related genes and the biofilm formation capacity were evaluated. A total of 77 Salmonella strains in 8 different serotypes were assessed: Salmonella Agona (n = 43), Salmonella Senftenberg (n = 13), Salmonella Meleagridis (n = 8), Salmonella Derby (n = 7), Salmonella Kottbus (n = 2), Salmonella Calabar (n = 2), Salmonella Kingston (n = 1), and Salmonella Typhimurium (n = 1). The results showed that all tested Salmonella strains produced biofilm at 25°C and 37°C after 3 d, and Salmonella Kingston and Salmonella Senftenberg had higher biofilm production than other strains under test conditions. Serotype, incubation temperature, pH, and their interactions had significant effects on biofilm formation for Salmonella. The strongest biofilm formation capacity of Salmonella (serovar Agona, Senftenberg, Kottbus, Calabar, Kingston, and Typhimurium) occurred at 25°C and at pH 7.0. Biofilm formation was significantly inhibited for all Salmonella strains incubated at 4°C. The detection rates of genes rpoS, fliC, wcaA, and invA were 100%, and the rates of genes csgB, csgD, csrA, sirA, adrA, gly, fimH, sdiA, ompR, sipB, sipC, luxS, and pfs exceeded 75% among all biofilm producer strains. The detection rate of igaA was significantly different between different biofilm producers. Based on the findings in this study, useful information on biofilm formation of Salmonella isolated from beef processing plants in China is provided, which could help clear the technological hurdle in delaying biofilm production to deal with risks from Salmonella biofilms in the beef industry.

Bio-protective potential of lactic acid bacteria: Effect of Lactobacillus sakei and Lactobacillus curvatus on changes of the microbial community in vacuum-packaged chilled beef.

OBJECTIVE: This study was to determine the bacterial diversity and monitor the community dynamic changes during storage of vacuum-packaged sliced raw beef as affected by Lactobacillus sakei and Lactobacillus curvatus. METHODS: L. sakei and L. curvatus were separately incubated in vacuumed-packaged raw beef as bio-protective cultures to inhibit the naturally contaminating microbial load. Dynamic changes of the microbial diversity of inoculated or non-inoculated (control) samples were monitored at 4°C for 0 to 38 days, using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). RESULTS: The DGGE profiles of DNA directly extracted from non-inoculated control samples highlighted the order of appearance of spoilage bacteria during storage, showing that Enterbacteriaceae and Pseudomonas fragi emerged early, then Brochothrix thermosphacta shared the dominant position, and finally, Pseudomonas putida showed up became predominant. Compared with control, the inoculation of either L. sakei or L. curvatus significantly lowered the complexity of microbial diversity and inhibited the growth of spoilage bacteria (p<0.05). Interestingly, we also found that the dominant position of L. curvatus was replaced by indigenous L. sakei after 13 d for L. curvatus-inoculated samples. Plate counts on selective agars further showed that inoculation with L. sakei or L. curvatus obviously reduced the viable counts of Enterbacteraceae, Pseudomonas spp. and B. thermosphacta during later storage (p< 0.05), with L. sakei exerting greater inhibitory effect. Inoculation with both bio-protective cultures also significantly decreased the total volatile basic nitrogen values of stored samples (p<0.05). CONCLUSION: Taken together, the results proved the benefits of inoculation with lactic acid bacteria especially L. sakei as a potential way to inhibit growth of spoilage-related bacteria and improve the shelf life of vacuum-packaged raw beef.

UPLC-Q-TOF/MS-based urine and plasma metabonomics study on the ameliorative effects of aspirin eugenol ester in hyperlipidemia rats.

The main objective of this study was to investigate the ameliorative effects of aspirin eugenol ester (AEE) in hyperlipidemic rat. After five-week oral administration of AEE in high fat diet (HFD)-induced hyperlipidemic rats, the impact of AEE on plasma and urine metabonomics was investigated to explore the underlying mechanism by UPLC-Q-TOF/MS analysis. Blood lipid levels and histopathological changes of liver, stomach and duodenum were also evaluated after AEE treatment. Without obvious gastrointestinal (GI) side effects, AEE significantly relieved fatty degeneration of liver and reduced triglyceride (TG), low density lipoprotein (LDL) and total cholesterol (TCH) (P<0.01). Clear separations of metabolic profiles were observed among control, model and AEE groups by using principal component analysis (PCA) and orthogonal partial least-squares-discriminate analysis (OPLS-DA). 16 endogenous metabolites in plasma and 18 endogenous metabolites in urine involved in glycerophospholipid metabolism, fatty acid metabolism, fatty acid beta-oxidation, amino acid metabolism, TCA cycle, sphingolipid metabolism, gut microflora and pyrimidine metabolism were considered as potential biomarkers of hyperlipidemia and be regulated by AEE administration. It might be concluded that AEE was a promising drug candidate for hyperlipidemia treatment. These findings could contribute to the understanding of action mechanisms of AEE and provide evidence for further studies.

Feces and liver tissue metabonomics studies on the regulatory effect of aspirin eugenol eater in hyperlipidemic rats.

BACKGROUND: Based on the pro-drug principle, aspirin and eugenol were esterified to synthesize aspirin eugenol ester (AEE). The anti-hyperlipidemia effect of aspirin eugenol ester has been confirmed in hyperlipidemic rat induced by high fat diet (HFD). However, its effect on liver and feces metabonomic profiles remains unknown. METHODS: Suspension of AEE was prepared in 5% carboxymethyl cellulose sodium (CMC-Na). Thirty rats were divided into control, model and AEE groups. The control and model rats were fed with normal diet or HFD for 13 weeks, respectively. Rats in AEE-treated group were fed with HFD for 8 weeks to induce hyperlipidemia, and then given AEE once daily by oral gavage for 5 weeks at the dosage of 54 mg/kg body weight. After drug intervention, lipid profile analysis and oil red O staining were carried out to confirm the lipid accumulation in liver tissue. UPLC-Q-TOF/MS-based liver and feces metabonomics coupled with pathway analysis were conducted to evaluate the changes of metabolic profile and endogenous metabolites. RESULTS: In liver tissue, oral administration of AEE significantly reduced lipid droplets and the levels of triglyceride (TG) and low-density lipoprotein (LDL). Using principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA), distinct changes in metabolite patterns in feces and liver were observed. Liver and feces samples in control, model and AEE groups were scattered in PLS-DA score plots. 28 metabolites in liver and 22 in feces were identified as potential biomarkers related to hyperlipidemia. As possible drug targets, the perturbations of those biomarkers can be regulated by administration of AEE. CONCLUSION: Anti-hyperlipidemia effect of AEE was confirmed by lipid analysis, oil red O staining and metabolomics analysis. The mechanism of AEE might be associated with the changes in the metabolism of glycerophospholipid, amino acid, fatty acid, sphingolipid, purine, bile acid and glutathione.

Synthesis and Biological Activity Evaluation of Novel Heterocyclic Pleuromutilin Derivatives.

A series of pleuromutilin derivatives were synthesized by two synthetic procedures under mild reaction conditions and characterized by Nuclear Magnetic Resonance (NMR), Infrared Spectroscopy (IR), and High Resolution Mass Spectrometer (HRMS). Most of the derivatives with heterocyclic groups at the C-14 side of pleuromutilin exhibited excellent in vitro antibacterial activities against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE), and vancomycin-resistant Enterococcus (VRE) in vitro antibacterial activity. The synthesized derivatives which contained pyrimidine rings, 3a, 3b, and 3f, displayed modest antibacterial activities. Compound 3a, the most active antibacterial agent, displayed rapid bactericidal activity and affected bacterial growth in the same manner as that of tiamulin fumarate. Moreover, molecular docking studies of 3a and lefamulin provided similar information about the interactions between the compounds and 50S ribosomal subunit. The results of the study show that pyrimidine rings should be considered in the drug design of pleuromutilin derivatives.